Our laboratory has recently discovered that ?E-catenin, a regulator of cell adhesion processes, plays a critical role in the development of the islet cell lineage by virtue of its function as a repressor of the Sonic Hedgehog (SHH) pathway. We found that deletion of ?E-catenin in Pdx1+ multipotent pancreatic progenitors results in the accumulation of immature bipotent Sox9+ progenitors. These ?E-cateninnull/Sox9+ progenitors are unable to adopt an endocrine cell phenotype due to a constitutive activation of the SHH pathway. Interestingly, pharmacological blockade of the SHH pathway in these ?E-cateninnull/Sox9+ progenitors restored their ability to differentiate into hormone expressing islet cells. More recently, we found that the temporal downregulation of ?E-catenin by siRNA in human adult islets can elicit significant ?-cell replication. Hence, based on these results, and on the notion that ?E-catenin can also block Wnt signaling, we hypothesize that in differentiated islet cells ?E-catenin may represent yet another ?brake? on cell cycle entry by virtue of its opposing functions on signaling SHH and Wnt pathways that would normally promote cell proliferation. To test this hypothesis, we will focus our studies on the following Specific Aims: Aim 1: Determine the role of ?E-catenin as a modulator of ?-cell growth during embryonic development and in postnatal life, under physiologic conditions and in injury settings. In these experiments will also test if the conditional deletion of ?E-catenin in embryonic and in postnatal ?-cells will de-repress SHH and Wnt, which in turn are expected to elicit cell cycle entry. In parallel studies we will also test if the conditional ablation of ?E-catenin in postnatal life will enhance ?-cell regeneration in the streptozotocin model of ?-cell injury, and/or in response to metabolic stressors such as exposure to high fat diet. Aim 2: Targeting ?E-catenin-dependent signaling axis for the ex vivo expansion of human islet cells and for the reprogramming of ductal cell populations, both in vitro and in vivo in cell transplantation models. Based on the notion that human islet cells exhibit a modest propensity to respond to pro-growth stimuli, these studies will test if knocking down ?E-catenin in human adult ?-cells will de-repress SHH and Wnt, and foster cell proliferation. A similar strategy will be tested on human adult ductal tissue, usually discarded from islet isolation procedures, to test if they can regain an embryonic-like competency to differentiate into endocrine cells, both in vitro and in vivo in cell transplantation models. Collectively, our strategy to transiently down-regulate ?E-catenin expression, allowing for the de-repression of the SHH pathway may prove as a powerful strategy to promote the ex vivo expansion of ?-cells, and/or re-direct the differentiation competency of adult ductal cells toward a ?-cell phenotype. Hence, we anticipate that these studies harbor significant translational value for cell replacement therapies in diabetes.